LOCUS       sequence-227514-        7840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  destination vector for GreenGate cloning method, contains 2x a set 
            of modules from A-F, ""Driver line"", tissues-specific promoter, 
            Dex-inducible mTurquoise2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7840)
  AUTHORS   Schurholz AK, Lopez-Salmeron V, Li Z, Forner J, Wenzl C, Gaillochet 
            C, Augustin S, Vilches-Barro A, Fuchs M, Gebert M, Lohmann JU, Greb 
            T, Wolf S
  TITLE     A Comprehensive Toolkit for Inducible, Cell Type-Specific Gene 
            Expression in Arabidopsis.
  JOURNAL   Plant Physiol. 2018 Jul 19. pii: pp.18.00463. doi: 
            10.1104/pp.18.00463.
  PUBMED    30026289
REFERENCE   2  (bases 1 to 7840)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant 
            Physiol. 2018 Jul 19. pii: pp.18.00463. doi: 10.1104/pp.18.00463."
COMMENT     Sequence Label: pSW505 - pOp4:3xGFP-NLS
FEATURES             Location/Qualifiers
     source          1..7840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    534..556
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     protein_bind    694..713
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    746..765
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    798..817
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    850..869
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     promoter        893..939
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             join(2524..2526,2527..2529,2530..3240)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2569..2590)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2830..2849)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3177..3198
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        4186..4566
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     misc_feature    5859..5883
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5974..6562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6054..6073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6733..7521)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      join(7812..7840,1..407)
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tttttatccc cggaagcctg tggatagagg gtagttatcc acgtgaaacc gctaatgccc
       61 cgcaaagcct tgattcacgg ggctttccgg cccgctccaa aaactatcca cgtgaaatcg
      121 ctaatcaggg tacgtgaaat cgctaatcgg agtacgtgaa atcgctaata aggtcacgtg
      181 aaatcgctaa tcaaaaaggc acgtgagaac gctaatagcc ctttcagatc aacagcttgc
      241 aaacacccct cgctccggca agtagttaca gcaagtagta tgttcaatta gcttttcaat
      301 tatgaatata tatatcaatt attggtcgcc cttggcttgt ggacaatgcg ctacgcgcac
      361 cggctccgcc cgtggacaac cgcaagcggt tgcccaccgt cgagcgcctt tgcccacaac
      421 ccggcggccg gccgcaacag atcgttttat aaattttttt ttttgaaaaa gaaaaagccc
      481 gaaaggcggc aacctctcgg gcttctggat ttccgatccc cggaattaga tcttggcagg
      541 atatattgtg gtgtaacgtt ctcgaactag gctcggacga agtaagcggc cgcaacctta
      601 attaaacacg gcgcgccaac agcgatcgca ccaggtacca cctatgcata tgtcgagctc
      661 aagaaatcta gaaagaagaa agggaagaga aagaattgtg agcgctcaca attgaaagac
      721 tagaaagaag aaagggaaga gaaagaattg tgagcgctca caattgaaag actagaaaga
      781 agaaagggaa gagaaagaat tgtgagcgct cacaattgaa agactagaaa gaagaaaggg
      841 aagagaaaga attgtgagcg ctcacaattg aaagactagt ggatcgatct tcgcaagacc
      901 cttcctctat ataaggaagt tcatttcatt tggagaggac gaacagtatt cagtcgactg
      961 gtaccaacag gctccatggt ttctaaaggt gaagagttgt ttactggagt tgtgcctatt
     1021 ctcgttgaac ttgatggaga tgtgaacgga cacaagttct ctgtttcagg agaaggagag
     1081 ggagatgcta cttatggaaa acttactttg aagtttattt gtactactgg aaaacttcct
     1141 gttccttggc ctactttggt gactactctt acttatggag tgcaatgctt ctcaagatac
     1201 ccagatcaca tgaagcagca cgatttcttc aaatctgcta tgcctgaagg ttatgttcaa
     1261 gagagaacaa ttttctttaa ggatgatgga aactacaaaa ctagagctga ggttaagttt
     1321 gagggagata cattggtgaa tagaattgaa ctcaagggaa tcgatttcaa agaggatggt
     1381 aacatccttg gacataaatt ggaatacaac tacaactcac acaacgttta catcatggct
     1441 gataagcaaa agaacggaat taaggtgaac ttcaagatca gacataacat cgaggatgga
     1501 tctgttcagt tggctgatca ctatcaacag aatactccaa ttggagatgg accagttctt
     1561 ttgcctgata accattacct ttctacacag tcaaagctgt ctaaagatcc taatgagaaa
     1621 agagatcaca tggttctcct tgagtttgtg actgctgcag gtatcacact cggtatggat
     1681 gagctttaca agggtacctc tggatctgga tcaggttctg gtagtggttc tggttcagga
     1741 tctgcatgca tggtttctaa gggtgaggaa ctcttcactg gtgttgttcc tatcctcgtt
     1801 gagcttgatg gtgatgtgaa cggacataag ttctctgttt ctggtgaagg tgagggtgat
     1861 gctacttacg gaaagcttac tctcaagttc atctgtacca ctggaaagtt gcctgttcct
     1921 tggcctactc tcgttactac tttcacttac ggtgtgcagt gcttctcaag ataccctgat
     1981 catatgaagc agcacgattt cttcaagtct gctatgcctg agggatacgt gcaagagaga
     2041 actatcttct tcaaggatga tggaaactac aagaccaggg ctgaggttaa gttcgagggt
     2101 gatactcttg tgaacaggat cgagcttaag ggaatcgatt tcaaagagga tggtaacatc
     2161 cttggacaca agcttgagta caactacaac tctcacaacg tgtacatcat ggctgataag
     2221 cagaagaacg gaatcaaggt taacttcaag atcaggcaca acatcgagga tggatctgtt
     2281 cagcttgctg atcattacca gcagaacact cctattggag atggacctgt tcttctccct
     2341 gataaccatt acctttctac ccagtctaag ctctctaaag atcctaacga gaagagggat
     2401 catatggtgc tcttggagtt tgttactgct gctggaatca ctcacggaat ggatgaactt
     2461 tacaagccgc ggtctggaag tggtagtgga agtggatcag gtagtggatc tggaagtacc
     2521 ggtatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     2581 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     2641 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     2701 accctcgtga ccaccttcac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     2761 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2821 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2881 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2941 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     3001 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     3061 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     3121 cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     3181 gtcctgctgg agttcgtgac cgccgccggg atcactcacg gcatggacga gctgtacaag
     3241 tcaggttctg gttcaggttc aggttctgga tctggaagtg gatctggatc agcggccgca
     3301 tctgaattca agcgtgaaga gcaagcaagg aaagctaagg tgaacaatga gaaaaagacg
     3361 gaaatagtga aaccagagag ttgtagcaat gaaggagatg tcaaggatct gaaaagaaag
     3421 gactctgagg atggaaacga gggtgaggaa gaagaagctt cttcgaaacc gaaaaagcca
     3481 aaagttgctc tttctcatct tcaggacatt gacgacacag aagctgatca agaagaagag
     3541 taactgcatc tcgtctctgt tatgcttaag aagttcaatg tttcgtttca tgtaaaactt
     3601 tggtggtttg tgttttgggg ccttgtataa tccctgatga ataagtgttc tactatgttt
     3661 ccgttcctgt tatctctttc tttctaatga caagtcgaac ttcttcttta tcatcgcttc
     3721 gtttttatta tctgtgcttc ttttgtttaa tacgcctgca aagtgactcg actctgttta
     3781 gtgcagttct gcgaaacttg taaatagtcc aattgttggc ctctagtaat agatgtagcg
     3841 aaagtgttga gctgttgggt tctaaggatg gcttgaacat gttaatcttt taggttctga
     3901 gtatgatgaa cattcgttgt tgctaagaaa tgcctgtaat gtcccacaaa tgtagaaaat
     3961 ggttcgtacc tttgtccaag cattgatatg tctgatgaga ggaaactgca agatactgag
     4021 cttggtttaa cgaaggagag gcagtttctt ccttccaaag catttcattt gacaatgcct
     4081 tgatcatctt aagtagagtt tctgttgtgg aaagtttgaa actttgaaga aacgactctc
     4141 aagtaaattg atgatcacaa gtgaaagtgt atgttactag gatccttttc aaatcagtgc
     4201 gcaagacgtg acgtaagtat ccgagtcagt ttttattttt ctactaattt ggtcgtttat
     4261 ttcggcgtgt aggacatggc aaccgggcct gaatttcgcg ggtattctgt ttctattcca
     4321 actttttctt gatccgcagc cattaacgac ttttgaatag atacgctgac acgccaagcc
     4381 tcgctagtca aaagtgtacc aaacaacgct ttacagcaag aacggaatgc gcgtgacgct
     4441 cgcggtgacg ccatttcgcc ttttcagaaa tggataaata gccttgcttc ctattatatc
     4501 ttcccaaatt accaatacat tacactagca tctgaatttc ataaccaatc tcgatacacc
     4561 aaatcgccat ggcttctatg atatcctctt cagctgtgac tacagtcagc cgtgcttcta
     4621 cggtgcaatc ggccgcggtg gctccattcg gcggcctcaa atccatgact ggattcccag
     4681 ttaagaaggt caacactgac attacttcca ttacaagcaa tggtggaaga gtaaagtgca
     4741 tggtgacggt gttcggcatt ctgaatctca ccgaggactc cttcttcgat gagagccggc
     4801 ggctagaccc cgccggcgct gtcaccgcgg cgatcgaaat gctgcgagtc ggatcagacg
     4861 tcgtggatgt cggaccggcc gccagccatc cggacgcgag gcctgtatcg ccggccgatg
     4921 agatcagacg tattgcgccg ctcttagacg ccctgtccga tcagatgcac cgtgtttcaa
     4981 tcgacagctt ccaaccggaa acccagcgct atgcgctcaa gcgcggcgtg ggctacctga
     5041 acgatatcca aggatttcct gaccctgcgc tctatcccga tattgctgag gcggactgca
     5101 ggctggtggt tatgcactca gcgcagcggg atggcatcgc cacccgcacc ggtcaccttc
     5161 gacccgaaga cgcgctcgac gagattgtgc ggttcttcga ggcgcgggtt tccgccttgc
     5221 gacggagcgg ggtcgctgcc gaccggctca tcctcgatcc ggggatggga tttttcttga
     5281 gccccgcacc ggaaacatcg ctgcacgtgc tgtcgaacct tcaaaagctg aagtcggcgt
     5341 tggggcttcc gctacttgtc tcggtgtcgc ggaaatcctt cttgggcgcc accgttggcc
     5401 ttcctgtaaa ggatctgggt ccagcgagcc ttgcggcgga acttcacgcg atcggcaatg
     5461 gcgctgacta cgtccgcacc cacgcgcctg gagatctgcg aagcgcaatc accatctcgg
     5521 aaaccctcgc gaaatttcgc agtcgcgacg ccagagacag agggttagat catgcctagc
     5581 ttggctgcag gtcgacctgc aggcatgccc gctgaaatca ccagtctctc tctacaaatc
     5641 tatctctctc tataataatg tgtgagtagt tcccagataa gggaattagg gttcttatag
     5701 ggtttcgctc atgtgttgag catataagaa acccttagta tgtatttgta tttgtaaaat
     5761 acttctatca ataaaatttc taattcctaa aaccaaaatc caggggtacc gtattctaga
     5821 ccaagcatgc acagctggcg ttctcagcca gatccccttg acaggatata ttggcgggta
     5881 aactaagtcg ctgtatgtgt ttgtttgaga tctcatgtga gcaaaaggcc agcaaaaggc
     5941 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6001 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6061 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6121 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6181 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6241 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6301 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6361 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6421 atttggtatc tgcgctctgc tgaagccagt taccttcgga agaagagttg gtagctcttg
     6481 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6541 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6601 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6661 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgtgtaaca
     6721 ttggtctagt gattatttgc cgactacctt ggtgatctcg cctttcacgt agtgaacaaa
     6781 ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tgtccaagat aagcctgcct
     6841 agcttcaagt atgacgggct gatactgggc cggcaggcgc tccattgccc agtcggcagc
     6901 gacatccttc ggcgcgattt tgccggttac tgcgctgtac caaatgcggg acaacgtaag
     6961 cactacattt cgctcatcgc cagcccagtc gggcggcgag ttccatagcg ttaaggtttc
     7021 atttagcgcc tcaaatagat cctgttcagg aaccggatca aagagttcct ccgccgctgg
     7081 acctaccaag gcaacgctat gttctcttgc ttttgtcagc aagatagcca gatcaatgtc
     7141 gatcgtggct ggctcgaaga tacctgcaag aatgtcattg cgctgccatt ctccaaattg
     7201 cagttcgcgc ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa caatggtgac
     7261 ttctacagcg cggagaatct cgctctctcc aggggaagcc gaagtttcca aaaggtcgtt
     7321 gatcaaagct cgccgcgttg tttcatcaag ccttacggtc accgtaacca gcaaatcaat
     7381 atcactgtgt ggcttcaggc cgccatccac tgcggagccg tacaaatgta cggccagcaa
     7441 cgtcggttcg agatggcgct cgatgacgcc aactacctct gatagttgag tcgatacttc
     7501 ggcgatcacc gcttccctca taacacccct tgtattactg tttatgtaag cagacagttt
     7561 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     7621 aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcaga tcacgcatct
     7681 tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca actggtccac
     7741 ctacaacaaa gctctcatca accgtggctc cctcactttc tggctggatg atggggcgat
     7801 tcaggcgatc cccatccaac agcccgccgt cgagcgggct
//
